Isolation and Screening of Laccase Producing Basidiomycetes via Submerged Fermentations
Approximately 10,000 different types of dyes and
pigments are being used in various industrial applications yearly,
which include the textile and printing industries. However, these dyes
are difficult to degrade naturally once they enter the aquatic system.
Their high persistency in natural environment poses a potential health
hazard to all form of life. Hence, there is a need for alternative dye
removal strategy in the environment via bioremediation. In this study,
fungi laccase is investigated via commercial agar dyes plates and
submerged fermentation to explore the application of fungi laccase in
textile dye wastewater treatment. Two locally isolated basidiomycetes
were screened for laccase activity using media added with commercial
dyes such as 2, 2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid
(ABTS), guaiacol and Remazol Brillant Blue R (RBBR). Isolate TBB3
(1.70±0.06) and EL2 (1.78±0.08) gave the highest results for ABTS
plates with the appearance of greenish halo on around the isolates.
Submerged fermentation performed on Isolate TBB3 with the
productivity 3.9067 U/ml/day, whereas the laccase activity for Isolate
EL2 was much lower (0.2097 U/ml/day). As isolate TBB3 showed
higher laccase production, it was subjected to molecular
characterization by DNA isolation, PCR amplification and sequencing
of ITS region of nuclear ribosomal DNA. After being compared with
other sequences in National Center for Biotechnology Information
(NCBI database), isolate TBB3 is probably from species Trametes
hirsutei. Further research work can be performed on this isolate by
upscale the production of laccase in order to meet the demands of the
requirement for higher enzyme titer for the bioremediation of textile
dyes.
[1] L., Mendoza, “Laccases from new fungal sources and some promising
applications”, PhD., Lunk University, 2011.
[2] H. B. Bhimani, “Bacterial degradation of azo dyes and its derivatives”,
PhD. Saurashtra University, 2011.
[3] Y. L. Pang, and A. Z. Abdullah, “Current status of textile industry
wastewater management and research progress in Malaysia: A Review”,
Clean-Soil, Air, Water, vol. 41, no. 8, pp. 751-764, 2012.
[4] D. Balan, and R. Monteiro, “Decolorization of textile indigo dye by
ligninolytic fungi”, J. of Biotech., vol. 89, pp. 141-145, 2001.
[5] G. Sumbali, “The Fungi”, 2nd ed., USA: Alpha Science Intl Ltd., 2011.
[6] F. Patrick, G. Mtui, A. M. Mshandete, and A. Kivaisi, “Optimization of
laccase and manganese peroxidase production in submerged culture of
Pleurotus sajorcaju”, African J. of Biotech., vol. 10, no. 50, pp. 10166-
10177, 2011.
[7] M. Narkhede, R. Mahajan, and K. Narkhede, “Ligninolytic enzyme
production and Remazol Brilliant Blue R (RBBR) decolorization by a
newly isolated white rot fungus: Basidiomycota spp. L-168.”, Int. J. of
Pharma and Bio Sciences, vol. 4, no. 1, pp. 220-228, 2013.
[8] A. Dhouib, M. Hamza, H. Zouari, T. Mechichi, R. H’midi, M. Labat, M.J.
Martínez, and S. Sayadi, Autochthonous fungal strains with high
ligninolytic activities from Tunisian biotopes, African J. of Biotech., vol.
4, no. 5, pp. 431-436, 2005.
[9] L.L. Kiiskinen, M. Ratto, and K. Kruus, Screening for novel laccaseproducing
microbes, J. of App.Microbiol., vol. 97, pp.640–646, 2004.
[10] G. R Jebapriya, and J. J. Gnanadoss, “Screening and molecular
characterization of white rot fungi capable of laccase production and dye decolourization”, Int. J. of Life Science & Pharma Research, vol. 4, no. 2,
pp. 12-20, 2014.
[11] M. Chander, and D. S. Arora, “Evaluation of some white-rot fungi for
their potential to decolourise industrial dyes”, Dyes and Pigments, vol. 72,
no. 2, pp. 192–198, 2007.
[12] D. Jing, “Improving the simultaneous production of laccase and lignin
peroxidase from Streptomyces lavendulae by medium optimization”,
Bioresource Tech., vol. 101, pp. 7592-7597, 2010.
[13] Y. Y Sim, “Molecular identification of Schizophyllum commune and its
production of Schizophyllan”, BSc. University Tunku Abdul Rahman,
2014.
[14] D. Lii, S. Coloe, R. Baird, and J. Pederson, “Rapid mini preparation of
fungal DNA for PCR”, J. of Clin. Microbiol., vol. 38, no. 1, pp. 471, 2000.
[15] G. M. Soares, M. T. de Amorim, and M. Costa-Ferreira, “Use of laccase
together with redox mediators to decolourize Remazol Brilliant Blue R”,
J. Biotechnol., vol. 89, pp. 123–129, 2001.
[16] F. Patrick, G. Mtui, A. M. Mshandete, and A. Kivaisi, “Optimized
production of lignin peroxidase, manganese peroxidase and laccase in
submerged cultures of Trametes trogii using various growth media
compositions”, Tanz. J. Sci., vol 36, pp 1-18, 2010.
[17] E. Kalmış, Í. Yaşa, F. Kalyoncu, B. Pazarbaşı, and A. Koçyiǧit,
“Ligninolytic enzyme activities in mycelium of some wild and
commercial mushrooms”, Afr. J. of Biochem. Research, vol. 7, no. 23, pp.
4313-4320, 2008.
[18] C. M. Maganhotto de Souza Silva, I. Soares de Melo, and P. Roberto de
Oliveira, “Ligninolytic enzyme production by Ganoderma spp.”, Enz.and
Microbiol. Tech., vol. 37, pp. 324-329, 2004.
[19] E. Gomes, A. P. Aguiar, C. C. Carvalho, M. R. B. Bonfá, R. D. Silva, and
M. Boscolo, “Ligninases production by basidiomycetes strains on
ligninocellulosic argicultural residues and their application in the
decolorization of synthetic dyes”, Brazilian J. of Microbiol., vol. 40, pp.
31-39, 2009.
[20] E. Erden, M. C. Ucar, Tekin Gezer, and N. K. Pazarlioglue, “Screening
for ligninolytic enzymes from autochthonous fungi and applications for
decolorization of remazole marine blue”, Brazilian J. of Microbiol, vol.
40, pp. 346-353, 2009.
[21] H. W. Heldt, “Plant Biochemistry”, 3rd ed., Elsevier Academic Press,
2005.
[22] E. M. Fox, and B. J. Howlett, “Secondary metabolism: regulation and role
in fungal biology”, Curr. Opinion in Microbiol., vol. 11, pp. 481-487,
2008.
[23] M. H. Gold, and H. Alic, “Molecular biology of the lignin-degrading
basidiomycete Phanerochaete chrysosporium”, Microbiol. Rev., vol. 57,
no. 3, pp. 605-622, 1993.
[24] C. Eggert, U. Temp, and K. E. L. Eriksson, “The ligninolytic system of
the white rot fungus Pynoporus cinnabarinus: purification and
characterization of the laccase”, App. and Env. Microbiol., vol. 62 no. 4,
pp. 1151-1158, 1995.
[1] L., Mendoza, “Laccases from new fungal sources and some promising
applications”, PhD., Lunk University, 2011.
[2] H. B. Bhimani, “Bacterial degradation of azo dyes and its derivatives”,
PhD. Saurashtra University, 2011.
[3] Y. L. Pang, and A. Z. Abdullah, “Current status of textile industry
wastewater management and research progress in Malaysia: A Review”,
Clean-Soil, Air, Water, vol. 41, no. 8, pp. 751-764, 2012.
[4] D. Balan, and R. Monteiro, “Decolorization of textile indigo dye by
ligninolytic fungi”, J. of Biotech., vol. 89, pp. 141-145, 2001.
[5] G. Sumbali, “The Fungi”, 2nd ed., USA: Alpha Science Intl Ltd., 2011.
[6] F. Patrick, G. Mtui, A. M. Mshandete, and A. Kivaisi, “Optimization of
laccase and manganese peroxidase production in submerged culture of
Pleurotus sajorcaju”, African J. of Biotech., vol. 10, no. 50, pp. 10166-
10177, 2011.
[7] M. Narkhede, R. Mahajan, and K. Narkhede, “Ligninolytic enzyme
production and Remazol Brilliant Blue R (RBBR) decolorization by a
newly isolated white rot fungus: Basidiomycota spp. L-168.”, Int. J. of
Pharma and Bio Sciences, vol. 4, no. 1, pp. 220-228, 2013.
[8] A. Dhouib, M. Hamza, H. Zouari, T. Mechichi, R. H’midi, M. Labat, M.J.
Martínez, and S. Sayadi, Autochthonous fungal strains with high
ligninolytic activities from Tunisian biotopes, African J. of Biotech., vol.
4, no. 5, pp. 431-436, 2005.
[9] L.L. Kiiskinen, M. Ratto, and K. Kruus, Screening for novel laccaseproducing
microbes, J. of App.Microbiol., vol. 97, pp.640–646, 2004.
[10] G. R Jebapriya, and J. J. Gnanadoss, “Screening and molecular
characterization of white rot fungi capable of laccase production and dye decolourization”, Int. J. of Life Science & Pharma Research, vol. 4, no. 2,
pp. 12-20, 2014.
[11] M. Chander, and D. S. Arora, “Evaluation of some white-rot fungi for
their potential to decolourise industrial dyes”, Dyes and Pigments, vol. 72,
no. 2, pp. 192–198, 2007.
[12] D. Jing, “Improving the simultaneous production of laccase and lignin
peroxidase from Streptomyces lavendulae by medium optimization”,
Bioresource Tech., vol. 101, pp. 7592-7597, 2010.
[13] Y. Y Sim, “Molecular identification of Schizophyllum commune and its
production of Schizophyllan”, BSc. University Tunku Abdul Rahman,
2014.
[14] D. Lii, S. Coloe, R. Baird, and J. Pederson, “Rapid mini preparation of
fungal DNA for PCR”, J. of Clin. Microbiol., vol. 38, no. 1, pp. 471, 2000.
[15] G. M. Soares, M. T. de Amorim, and M. Costa-Ferreira, “Use of laccase
together with redox mediators to decolourize Remazol Brilliant Blue R”,
J. Biotechnol., vol. 89, pp. 123–129, 2001.
[16] F. Patrick, G. Mtui, A. M. Mshandete, and A. Kivaisi, “Optimized
production of lignin peroxidase, manganese peroxidase and laccase in
submerged cultures of Trametes trogii using various growth media
compositions”, Tanz. J. Sci., vol 36, pp 1-18, 2010.
[17] E. Kalmış, Í. Yaşa, F. Kalyoncu, B. Pazarbaşı, and A. Koçyiǧit,
“Ligninolytic enzyme activities in mycelium of some wild and
commercial mushrooms”, Afr. J. of Biochem. Research, vol. 7, no. 23, pp.
4313-4320, 2008.
[18] C. M. Maganhotto de Souza Silva, I. Soares de Melo, and P. Roberto de
Oliveira, “Ligninolytic enzyme production by Ganoderma spp.”, Enz.and
Microbiol. Tech., vol. 37, pp. 324-329, 2004.
[19] E. Gomes, A. P. Aguiar, C. C. Carvalho, M. R. B. Bonfá, R. D. Silva, and
M. Boscolo, “Ligninases production by basidiomycetes strains on
ligninocellulosic argicultural residues and their application in the
decolorization of synthetic dyes”, Brazilian J. of Microbiol., vol. 40, pp.
31-39, 2009.
[20] E. Erden, M. C. Ucar, Tekin Gezer, and N. K. Pazarlioglue, “Screening
for ligninolytic enzymes from autochthonous fungi and applications for
decolorization of remazole marine blue”, Brazilian J. of Microbiol, vol.
40, pp. 346-353, 2009.
[21] H. W. Heldt, “Plant Biochemistry”, 3rd ed., Elsevier Academic Press,
2005.
[22] E. M. Fox, and B. J. Howlett, “Secondary metabolism: regulation and role
in fungal biology”, Curr. Opinion in Microbiol., vol. 11, pp. 481-487,
2008.
[23] M. H. Gold, and H. Alic, “Molecular biology of the lignin-degrading
basidiomycete Phanerochaete chrysosporium”, Microbiol. Rev., vol. 57,
no. 3, pp. 605-622, 1993.
[24] C. Eggert, U. Temp, and K. E. L. Eriksson, “The ligninolytic system of
the white rot fungus Pynoporus cinnabarinus: purification and
characterization of the laccase”, App. and Env. Microbiol., vol. 62 no. 4,
pp. 1151-1158, 1995.
@article{"International Journal of Biological, Life and Agricultural Sciences:72063", author = "Mun Yee Chan and Sin Ming Goh and Lisa Gaik Ai Ong", title = "Isolation and Screening of Laccase Producing Basidiomycetes via Submerged Fermentations", abstract = "Approximately 10,000 different types of dyes and
pigments are being used in various industrial applications yearly,
which include the textile and printing industries. However, these dyes
are difficult to degrade naturally once they enter the aquatic system.
Their high persistency in natural environment poses a potential health
hazard to all form of life. Hence, there is a need for alternative dye
removal strategy in the environment via bioremediation. In this study,
fungi laccase is investigated via commercial agar dyes plates and
submerged fermentation to explore the application of fungi laccase in
textile dye wastewater treatment. Two locally isolated basidiomycetes
were screened for laccase activity using media added with commercial
dyes such as 2, 2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid
(ABTS), guaiacol and Remazol Brillant Blue R (RBBR). Isolate TBB3
(1.70±0.06) and EL2 (1.78±0.08) gave the highest results for ABTS
plates with the appearance of greenish halo on around the isolates.
Submerged fermentation performed on Isolate TBB3 with the
productivity 3.9067 U/ml/day, whereas the laccase activity for Isolate
EL2 was much lower (0.2097 U/ml/day). As isolate TBB3 showed
higher laccase production, it was subjected to molecular
characterization by DNA isolation, PCR amplification and sequencing
of ITS region of nuclear ribosomal DNA. After being compared with
other sequences in National Center for Biotechnology Information
(NCBI database), isolate TBB3 is probably from species Trametes
hirsutei. Further research work can be performed on this isolate by
upscale the production of laccase in order to meet the demands of the
requirement for higher enzyme titer for the bioremediation of textile
dyes.", keywords = "Bioremediation, dyes, fermentation, laccase.", volume = "10", number = "2", pages = "77-4", }
